title: Sarcoplasmic reticulum Ca2+-ATPase from rabbit skeletal muscle modified by peroxynitrite.
abstract: OBJECTIVE
  Effect of peroxynitrite on SERCA1 activity was studied in correlation with enzyme carbonylation. Kinetic parameters and location of peroxynitrite effect on SERCA1 were determined.
  
  
  METHODS
  Carbonyls were determined by immunoblotting. FITC, NCD-4 and Trp fluorescence were used to indicate changes in cytosolic and transmembrane regions of SERCA1.
  
  
  RESULTS
  Peroxynitrite-concentration-dependent decrease of SERCA1 activity was associated with elevation of protein carbonyls. 4-HNE was not involved in carbonylation of SERCA1. Increased FITC fluorescence in the presence of peroxynitrite correlated with the decrease of the enzyme affinity to ATP.
  
  
  CONCLUSIONS
  Peroxynitrite-induced SERCA1 carbonylation that was not accompanied with the formation of 4-HNE-SERCA1 adducts is indicative of direct oxidation of SERCA1. As assessed by FITC fluorescence and decreased affinity of the enzyme to ATP, peroxynitrite impairment was found to occur in the cytosolic ATP-binding region of SERCA1.
authors.affiliation: Institute of Experimental Pharmacology and Toxicology, Slovak Academy of Sciences , Bratislava , Slovak Republic .
authors.forename: Petronela
authors.initials: P
authors.lastname: Žižková
authors.forename: Jana
authors.initials: J
authors.lastname: Viskupičová
authors.forename: Dušan
authors.initials: D
authors.lastname: Blaškovič
authors.forename: Miriam K
authors.initials: MK
authors.lastname: Štrosová
authors.forename: Neven
authors.initials: N
authors.lastname: Žarkovič
authors.forename: Lubica
authors.initials: L
authors.lastname: Horáková
mesh_terms: D000818:Animals; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D015394:Molecular Structure; D018482:Muscle, Skeletal / Q000201:enzymology* / Q000378:metabolism; D030421:Peroxynitrous Acid / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D011817:Rabbits; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D004791:Enzyme Inhibitors; D030421:Peroxynitrous Acid; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases
keywords: 4-Hydroxynonenal; calcium pump; carbonyls; oxidative stress